Zinc oxide promotes poultry growth, but it tends to agglomerate. This necessitates high doses and leads to environmental contamination from unabsorbed, excreted zinc. Undigested zinc is excreted and can enter the food chain, increasing the probability of zinc residues in edible poultry tissues (muscle, liver, and eggs) and raising concerns for consumer safety. MOF-supported single-atom zinc catalysts (SAC) resolve agglomeration by atomic anchoring, enhancing bioavailability. High-temperature/high-pressure fixation of Zn2+ surfaces was confirmed by XRD, while FESEM revealed the corresponding surface morphology, collectively verifying SAC formation. SAC exhibited potent antimicrobial efficacy against key pathogens such as Salmonella typhimurium, Escherichia coli, and Staphylococcus aureus (MIC of 3.125 mg/mL, MBC of 25 mg/mL). Co-culture experiments further demonstrated that the antibacterial performance of SAC remained stable over a temperature range of 20–80 °C and a pH range of 2–8, thus exhibiting excellent thermal stability and gastrointestinal tolerance. In 7-day-old chicks, SAC alleviated S. typhimurium-induced inflammation, reduced bacterial adherence, upregulated claudin-1, preserved gut homeostasis, ameliorated tissue lesions, and increased the abundance of Lactobacillus in the cecum, demonstrating promising potential for poultry infection control.
Lactobacilli species have emerged as a focal point in food microbiology due to their core probiotic properties, including the regulation of intestinal homeostasis and the enhancement of immunity. This study focuses on Lacticaseibacillus rhamnosus MG0718 (hereinafter referred to as MG0718), employing a combined approach of phenotypic evaluation and whole-genome sequencing to assess its probiotic potential and analyze the correlation between its phenotype and genotype. In vitro experiments demonstrated that MG0718 possesses broad-spectrum antibacterial activity against pathogenic bacteria. In vitro experiments showed that MG0718 had broad-spectrum antibacterial activity against pathogenic bacteria such as Escherichia coli (E. coli), with an inhibition zone diameter of up to 13.67 ± 1.56 mm. It survived pH 2.5 for 6 h with only a 1.72 log10 reduction, and showed 0.78 and 1.11 log10 CFU/mL reductions in artificial gastric and intestinal fluids after 2 h. DPPH scavenging was 56.7% and total reducing power was 91.1%. In vivo, 7-day preventive administration maintained 100% survival against S. Typhimurium infection and alleviated weight loss. Bacterial loads in spleen, liver, and cecum dropped from 4.5, 4.5, and 4.2 to 3.6, 1.8, and 2.5 lg CFU/g, respectively. Whole-genome sequencing analysis indicated that the complete genome of MG0718 is 2,574,565 bp in length, containing 2813 CDS. Among these genomic components, 203 stress-related protein genes elucidate its superior environmental tolerance; one bacteriocin gene cluster, one EPS gene cluster and two secondary metabolite gene clusters provide the genetic basis for its antibacterial activity. Notably, no virulence factors were detected, ensuring the safety of the strain for application. In summary, the functional phenotypes of MG0718 are highly consistent with its genetic characteristics, identifying it as a probiotic candidate of significant developmental value. Future research should focus on clinical trials to further verify its practical benefits for human intestinal health and immunomodulation, thereby providing a robust scientific basis for its application in functional foods.
The experiment aims to optimize the extraction process of polysaccharides and coumarins from Ficus hirta Vahl by water bath and to clarify its in vitro antioxidant activity. Using dry powder yield, polysaccharide content, and coumarin content as indicators, the effects of liquid to material ratio, extraction time, extraction temperature, and extraction times on the extraction efficiency of polysaccharides and coumarins were investigated through single factor experiments, orthogonal experiments, and multi index comprehensive scoring methods. The results showed that the optimal process for extracting polysaccharides and coumarins of Ficus hirta Vahl using water bath extraction method was a liquid to material ratio of 20 mL/g, extraction time of 60 min, extraction temperature of 80 ℃, and extraction frequency of 2 times. Under these conditions, the polysaccharide content of the Ficus hirta Vahl was 59.50% and and coumarin content was 2.28%. The Ficus hirta Vahl extract exhibited good scavenging effects on 1, 1-diphenyl-2-trinitrophenylhydrazine (DPPH) radical and hydroxyl radical. The study shows that this extraction method is stable, easy to operate, environmentally friendly, and highly reproducible, providing a reference for the further development and utilization of Ficus hirta Vahl in the fields of feed and agriculture.
This study investigated the effects of composite postbiotic preparation on the growth performance, immune function, and microbiota composition of Nubian black goats. Thirty healthy Nubian black goats with similar body weights were randomly assigned to two groups (n = 15 per group): a control group fed a basal diet and a treatment group fed the basal diet supplemented with 0.5% composite postbiotic preparation (equal-ratio co-fermentation of Bacillus subtilis GX15 and Lentilactobacillus buchneri GX0328-6). The results indicated that while compound postbiotic supplementation did not significantly alter the average daily gain (ADG) and the serum biochemical indices (p > 0.05), it significantly increased the concentrations of immunoglobulins (IgG and IgA) compared to the control group (p < 0.05). Despite comparable α- and β-diversity, CPP supplementation selectively enriched Bacteroides, UCG-005, and Ruminococcaceae while reducing Turicibacter (LEfSe LDA > 2.0; STAMP p < 0.05), suggesting targeted modulation of gut microbiota. In conclusion, dietary supplementation with 0.5% composite postbiotic preparation improves immune function and modulates intestinal microbiota composition without significantly affecting growth performance in black goats while improving intestinal microbial composition and promoting overall gut health.
An outbreak characterized by clinical signs of diarrhea and paralysis, occasionally progressing to fatal outcomes, occurred at an ostrich breeding facility. Conventional antibiotic treatments proved ineffective. To investigate the etiology of the disease, brain and liver specimens were collected for diagnostic analysis. An Escherichia coli (E. coli) isolate, designated strain HZDC01, was obtained from cerebral tissues, and whole-genome sequencing was performed for genomic characterization. Genomic analysis revealed that the chromosomal DNA harbors numerous resistance genes, conferring multidrug resistance through complex mechanisms. Furthermore, a p0111-type plasmid carrying the blaCTX-M-55 gene and an IncX1-type plasmid harboring rmtB, sul1, APH(6)-Id, tet(A), AAC(3)-IIc, aadA2, blaTEM-1B, and floR genes were identified. These plasmids carry numerous mobile genetic elements that can disseminate via horizontal gene transfer, thereby amplifying the risk of resistance-gene spread within bacterial populations. Additionally, the ibeB and ibeC genes, which encode proteins involved in the invasion of brain microvascular endothelial cells, were identified. These genes may facilitate E. coli penetration of the blood–brain barrier, potentially leading to meningitis and posing a life-threatening risk to the host. This is the first report of the isolation and characterization of extended-spectrum beta-lactamase E. coli from the brain of an ostrich with paralysis. The findings provide valuable genomic insights into the antimicrobial resistance profiles and pathogenic mechanisms of ostrich-derived E. coli isolates.
Lactobacilli, recognized as beneficial bacteria within the human body, are celebrated for their multifaceted probiotic functions, including the regulation of intestinal flora, enhancement of body immunity, and promotion of nutrient absorption. This study comprehensively analyzed the genotypic and phenotypic characteristics of Lactiplantibacillus plantarum (L. plantarum) strains isolated from the intestines of healthy chicks and assessed their potential as probiotics. The assembled genome consists of 29,521,986 bp, and a total of 1,771 coding sequences (CDSs) were predicted. Based on the entire genome sequence analysis, 50 stress resistance genes and seven virulence factors were identified. The results of the phenotypic experiments showed that the strain had good resistance to high temperature, low temperature, acid, alkali, salt, artificial gastrointestinal fluid, and strong antioxidant capacity. Additionally, transcriptomic analysis confirmed that under stress conditions, the expression levels of key genes were significantly upregulated. Therefore, the phenotypic characteristics of L. plantarum GX17 align well with its genotypic features, demonstrating promising probiotic properties. This strain holds great potential as a probiotic candidate, and further investigation into its beneficial effects on human health is warranted. IMPORTANCE:In humans, Lactiplantibacillus plantarum may synergize with host microbiota to ameliorate dysbiosis-related pathologies, enhance immunomodulation, and facilitate micronutrient bioavailability. For livestock, its application could improve feed conversion ratios, suppress enteric pathogens through competitive exclusion, and mitigate antibiotic overuse, "a critical strategy in One Health frameworks." Further investigations into strain-specific mechanisms (e.g., postbiotic metabolites, quorum sensing regulation) are warranted to translate these genomic-phenotypic advantages into sustainable health solutions across species.
Antibiotic-resistant bacteria are major contributors to food spoilage, animal diseases, and the emergence of multidrug-resistant (MDR) bacteria in healthcare, highlighting the urgent need for effective treatments. Bacteriocins produced by lactic acid bacteria (LAB) have gained attention for their non-toxic nature and strong antimicrobial properties. LAB-derived bacteriocins have been successfully applied in food preservation and are classified by the U.S. Food and Drug Administration (FDA) as ‘food-grade’ or ‘generally recognized as safe’ (GRAS). This review summarizes recent progress in the production, purification, and emerging applications of LAB bacteriocins. It emphasizes their versatility in food preservation, agriculture, and medicine, providing insights into their role in antimicrobial development and functional food innovation.
The escalating issue of bacterial resistance, coupled with stringent restrictions on antibiotic use in many countries, has prompted the search for alternative strategies to combat bacterial infections. Probiotics, such as Bacillus subtilis (B. subtilis), have been widely recognized for their ability to inhibit pathogenic bacteria proliferation and protect hosts from infection-related damage. In this study, a novel strain of B. subtilis, designated GX15, was isolated from the intestine of a healthy chicken. We systematically evaluated its in vitro antimicrobial activity and protective effects against Salmonella Typhimurium SM022 (S. Typhimurium SM022) infection in mice. GX15 exhibited broad-spectrum antibacterial activity, effectively inhibited the growth of both Gram-positive and Gram-negative bacteria, including S. Typhimurium SM022, Escherichia coli, and Staphylococcus aureus. In a murine model, S. Typhimurium SM022 infection induced clinical symptoms such as diarrhea, anorexia, and fur ruffling in C57BL/6 mice. Oral administration of GX15 at 10⁸ CFU/mL significantly attenuated these effects, reducing the elevation of immune organ indices, limiting bacterial translocation to peripheral tissues, and alleviating histopathological damage to the liver and intestinal tissues. Notably, GX15 did not alter antioxidant indices or immunoglobulin levels. However, it markedly modulated the expression of cytokines including GM-CSF, IL-6, TLR2, IL-10, and TGF-β in the small intestine, suggesting an immunoregulatory mechanism of action. B. subtilis GX15 exhibits potent probiotic activity and represents a promising prophylactic candidate against S. Typhimurium infections. These findings provide valuable insights into the potential use of GX15 as an alternative strategy for the prevention and control of salmonellosis.
The global epidemic of bovine coronavirus (BCoV) has caused enormous economic losses. The characterisation and genetic composition of endemic strains in Southwest China remain elusive. This study aimed to fill this gap by isolating three BCoV strains from this region and sequencing their whole genomes. To elucidate the genetic evolution and characterisation of the prevalent strains, the results of BCoV sequences were compared in GenBank, with a focus on genetic evolution, mutation, and recombination patterns. The results showed close homology between strains NN190313 and NN230328, while strain NN221214 showed less similarity to these two strains but clustered with the French strain of the European branch. Intriguingly, NN190313 and NN230328 were grouped with goat-derived BCoV strains from Jiangsu Province in Eastern China in the Asian–American branch. In addition, recombination analyses revealed significant signals between NN230328 and either a Chinese goat-derived strain (XJCJ2301G) or a Shandong strain (ShX310). This study highlights the importance of monitoring cross-species transmission between cattle and goats, especially in the mountainous areas of Southwest China where mixed farming occurs, and thus, the monitoring of cross-species transmission between cattle and goats is important for preventing new public health challenges, providing important insights for research on cross-species transmission, early prevention, and control measures, with potential applications in vaccine development.
The rising incidence of antibiotic resistance in Salmonella typhimurium indicates the urgent need for alternative antimicrobial agents. This study aims to identify natural compounds with antibacterial activity against S. typhimurium by screening the fermentation supernatant of Lactobacillus buchneri GX0328-6. Bacteriocins present in the supernatant were fractionated through ultrafiltration using membranes with molecular weight cut-offs of 1, 3, and 10 kDa. Further purification was achieved via gel filtration and reversed-phase high-performance liquid chromatography (RP-HPLC). Through this method, a novel bacteriocin was isolated and designated as GX-6. Its antimicrobial activity was assessed using the Oxford cup assay after neutralizing the effects of acid and hydrogen peroxide. GX-6 exhibited strong heat resistance, stability within a narrow pH range (pH 2.0-8.0), and a favorable safety profile. At a minimum inhibitory concentration (MIC) of 2 mg/mL, GX-6 effectively eradicated 99.9 % of S. typhimurium within 24 h (p < 0.0001). Scanning electron microscopy revealed that GX-6 disrupted bacterial cell membrane integrity, leading to surface crumpling, intracellular content leakage, and cell rupture. Thus, the novel bacteriocin, GX-6 demonstrates promising potential as an alternative therapeutic agent for S. typhimurium infectious disease. The main limitation of this work lies on the absence of in vivo evaluation of GX-6's antimicrobial activity and the lack of established large-scale industrial production methodologies, which should be fully explored before its widely applications. Thus, the novel bacteriocin, GX-6 demonstrates promising potential as an alternative therapeutic agent for S. typhimurium infectious disease.
Lactiplantibacillus plantarum strains are potentially rich sources of probiotics that could help avoid infections. In order to evaluate their efficacy in bolstering resistance to Salmonella typhimurium infection among chicks. In this study, L. plantarum and commercial probiotics were administered via the water supply at a dosage of 1×109 CFU per chicken from days 1 to 7 to establish a protective system for the chicks. On days 8 and 9, S. typhimurium was attacked to investigate the preventive effects and potential mechanisms of L. plantarum in comparison with commercial probiotics. Post-treatment, we took a broad range of measurements, including body weight, immune organ index changes, the viable count of S. typhimurium in the liver, spleen, and cecum, as well as pathological changes in the liver. Our findings demonstrated that both L. plantarum and the commercial probiotic could safeguard chicks from S. typhimurium infection. The data also suggested that probiotic medication could ease weight loss postinfection, lower the bacterial count in the liver, spleen, and cecum, and attenuate liver pathological damage among all treated participants. Subsequently, we did high-throughput sequencing of 16S rRNA to examine the fecal microbiota of the chicks 5 days post-infection. We discovered that both L. plantarum and the commercial probiotic could fend off the invasion of S. typhimurium by affecting the bacterial population of Anaerotruncus, Colidextribacter, and Lactobacillus. Generally speaking, the addition of L. plantarum as a feed additive protects yellow-feathered broilers from S. typhimurium illness, suggesting great potential for commercial uses in the poultry industry.
Salmonella Typhimurium (S. Typhimurium) contamination poses a significant challenge to breeder egg hatchability and chick health, necessitating the exploration of alternative disinfection methods. This study investigates the potential of phage vB_SPuM_SP02 (SP02) as a novel disinfectant for breeder eggs contaminated with S. Typhimurium SM022. Phage SP02 was isolated from poultry farm effluent and characterized for morphology, biological properties, and genome properties. Experimental groups of specific pathogen-free (SPF) eggs were treated with Salmonella and phage SP02, and efficacy was assessed through hatching rates, chick survival, weight, Salmonella load, immune organ indices, and intestinal flora. Phage treatment effectively eradicated Salmonella contamination on eggshells within 12 h, resulting in increased hatching and survival rates compared to controls. Furthermore, phage treatment mitigated weight loss and tissue Salmonella load in chicks without causing immune organ damage while reducing Salmonella spp. abundance in the intestinal tract. This study demonstrates the potential of phage SP02 as an eco-friendly and efficient disinfectant for S. Typhimurium-contaminated breeder eggs, offering promising prospects for practical application in poultry production.
Bovine coronavirus (BCoV) is a notable pathogen affecting newly born calves and adult cattle, increasing mortality rates among calves and reducing productivity in meat and dairy industries, thereby causing substantial economic losses. Current primary laboratory methods for detecting BCoV include RT-PCR assay, real-time RT-PCR assay, and ELISA. However, these methods are time-consuming, require specialized technicians, and necessitate a laboratory environment. Consequently, there is an urgent need for a rapid, sensitive, and easy to use diagnostic method to detect BCoV. This study introduces two innovative protocols: the real-time fluorescent reverse transcription recombinase-aided amplification (RT-RAA) and the test strip RT-RAA (RT-RAA-LFD). Our results indicate that real-time RT-RAA can complete the reaction in 20 min at 39°C, while RT-RAA-LFD can achieve detection in just 17.5 min at 35°C. These new approaches offer higher specificity, with no cross-reactivity to other viruses, and significantly enhanced sensitivity compared to existing methods (1.46 × 101 and 1.46 × 102 copies/μL, respectively). We evaluated the performance of our methods using 242 clinical samples, and compared with RT-PCR and RT-qPCR. Both real-time RT-RAA and RT-qPCR yielded similar detection rates, the detection rate of RT-RAA-LFD was better than RT-PCR. The RT-RAA methods developed in this study effectively overcome the limitations associated with both RT-PCR and RT-qPCR by offering advantages including a single, low reaction temperature that allows for room temperature operation. Both methods boast shorter reaction times, simpler and more portable instrumentation, as well as reduced technical and environmental demands. Generally, both RT-RAA methods established in this study offer new avenues for the rapid detection of BCoV, contributing significantly to the monitoring, prevention, and control of the disease in global bovine industry.
魏氏梭菌(Clostridium welchii),又称产气荚膜梭菌(Clostridium perfringens),菌体呈直杆状,革兰氏染色阳性,不具运动性.芽胞大而圆,位于菌体中央或近端,大部分菌株能产生荚膜.
Salmonellosis is a disease caused by non-typhoid Salmonella, and although some lactic acid bacteria strains have been shown previously to relieve Salmonellosis symptoms, little has been studied about the preventive mechanism of Lentilactobacillus buchneri (L. buchneri) against Salmonella infection in vivo. Therefore, the L. buchneri was fed to C57BL/6 mice for 10 days to build a protective system of mice to study its prevention and possible mechanisms. The results showed that L. buchneri GX0328-6 alleviated symptoms caused by Salmonella typhimurium infection among C57BL/6 mice, including low survival rate, weight loss, increase in immune organ index and hepatosplenomegaly, and modulated serum immunoglobulin levels and intrinsic immunity. Importantly, the L. buchneri GX0328-6 enhanced the mucosal barrier of the mouse jejunum by upregulating the expression of tight junction proteins such as ZO-1, occludins, and claudins-4 and improved absorptive capacity by increasing the length of mouse jejunal villus and the ratio of villus length to crypt depth and decreasing the crypt depth. L. buchneri GX0328-6 reduced the intestinal proliferation and invasion of Salmonella typhimurium by modulating the expression of antimicrobial peptides in the intestinal tract of mice, and reduced intestinal inflammation and systemic spread in mice by downregulating the expression of IL-6 and promoting the expression of IL-10. Furthermore, L. buchneri GX0328-6 increased the relative abundance of beneficial bacteria colonies and decreased the relative abundance of harmful bacteria in the cecum microflora by modulating the microflora in the cecum contents.
为了研究不同来源的地衣芽孢杆菌对常见致病菌的抑菌效果等生物学特性,试验采用细菌形态学、生理生化、16S rRNA基因测序、系统发育树分析、牛津杯抑菌试验等检测技术,对5株不同来源(鸡粪、羊粪、牛瘤胃内容物、猪肠内容物及土壤)的地衣芽孢杆菌进行了分离鉴定及体外抑菌活性检测.结果表明:5株分离株B17(鸡粪)、B30(羊粪)、B32(牛瘤胃内容物)、B35(猪肠内容物)和B38(土壤)均可在LB固体培养基上生长,其中有3株不同来源的分离株B30、B35和B38菌落形态相同;镜检可见分离株均为革兰氏阳性杆菌、均有近中生的孢子,符合芽孢杆菌的菌体特征;生理生化特性与芽孢杆菌相符;PCR扩增均得到约为1 500 bp的目的片段;所有分离株与GenBank中的地衣芽孢杆菌在同一簇,均鉴定为地衣芽孢杆菌;在5株地衣芽孢杆菌中,除猪源地衣芽孢杆菌B35外,其余4株均有抑菌作用,羊源及牛源地衣芽孢杆菌B30及B32的抑菌效果较好,抑菌范围更广泛.说明从羊粪及牛瘤胃内容物中分离出的地衣芽孢杆菌可作为益生芽孢杆菌控制病原菌,不同来源的地衣芽孢杆菌的抑菌效果不同.
Lactobacillus plantarum has recently been found to be a natural source feed additive bacteria with great advantages in food safety and animal welfare. Discovering novel strains with commercial application potentiation could benefit the local poultry industry, and in particular support Chinese farmers. In this study, we tested a recently isolated novel strain of Lactobacillus plantarum GX17 as a feed additive on the growth performance and intestinal barrier functions of 1-day-old Chinese yellow-feather chicks. As good as other commercial probiotics, feeding with Lactobacillus plantarum GX17 showed significant improvements in humoral immune responses and enhanced the immune effect after vaccination for either the Newcastle disease vaccine or the avian influenza vaccine. This study also found that feeding with Lactobacillus plantarum GX17 improved the feed-to-weight ratio and caused a significant increase of the villus length to crypt depth ratio. Furthermore, Lactobacillus plantarum GX17 significantly up-regulated the mRNA expression of CLDN, MUC2, and TLR2, all of which are jejunum-associated barrier genes, indicating an improvement of the intestinal barrier functions by enhancing the tight junction between epithelia cells. These results are comparable to the effects of feeding the commercial complex probiotics that improve the expression levels of CLDN, ocludin, MUC2, TLR2, and TLR4. In terms of maintaining intestinal health, commercial complex probiotics increased the relative abundance of Parabacteroides and Romboutsia, while Lactobacillus plantarum GX17 increased the relative abundance of Pseudoflavonifractor. Our data suggest that Lactobacillus plantarum GX17 could enhance the intestinal absorption of nutrients and therefore improve the growth performance of Chinese yellow-feather chicks. In conclusion, compared with the commercial complex probiotics, Lactobacillus plantarum GX17 has more positive effects on the growth performance and intestinal barrier function of yellow-feather chickens, and can be used as a feed additive.
[目的]试验旨在研究植物乳杆菌GX20200417-1菌株的生物学特性及体外抑菌活性,探讨其在生产中应用的可能性.[方法]将植物乳杆菌GX20200417-1菌株接种至不同pH和含不同浓度胆盐的PBS,以及人工胃肠液中,使用菌落计数分析其对酸、胆盐和人工胃肠液的耐受性.使用牛津杯法检测GX20200417-1菌株对大肠杆菌、金黄色葡萄球菌及沙门氏菌的抑菌能力;通过与霉菌毒素共培养后使用ELISA方法研究GX20200417-1菌株对霉菌毒素的降解能力,并饲喂小鼠进行安全性试验.[结果]植物乳杆菌GX20200417-1菌株能够耐受pH 2.0及0.3%胆盐,并在人工胃肠液中有至少1×104 CFU/mL的活菌数;GX20200417-1菌株发酵上清液对大肠杆菌、沙门氏菌、金黄色葡萄球菌均有明显的抑制作用,对玉米赤霉素及黄曲霉毒素均有降解作用,但对呕吐毒素无降解作用;灌服GX20200417-1菌株后小鼠安全、无毒副作用.[结论]植物乳杆菌GX20200417-1菌株具有优良的益生特性,在畜禽生产中有一定的开发潜力.
近年来,畜禽规模化养殖蓬勃发展,但我国畜禽养殖业标准化工作相对滞后,而畜禽养殖业的标准化建设是畜禽养殖业发展方式转型升级的必由之路.只有加快我国畜禽养殖业标准化工作,才能促进畜禽养殖业产业化、品牌化发展,助力乡村振兴.本文分析加快我国畜禽养殖业标准化工作的重要意义,并提出相应的解决方法.
Abstract To investigate effects of Lactobacillus plantarum and Bacillus subtilis as feed additives on the growth performance of near maturity yellow-feathered broilers. A total of 75 100-day-old hens were randomly divided into five groups. 1% of L.plantarum(P-group), B.subtilis (B-group), a 1:1 mixture of both strains(PB-group), or commercial probiotic production(M-group)were added into the drinking water at a concentration of 1.5x109 cfu/mL respectively. The control group was fed basal diet. Our results showed that L.plantarum could increase the intestinal enzyme activity of broilers, while decreasing the plasma MDA content, increasing the ALB and TP content, decreasing the plasma AST content, increasing the concentrations of plasma IgA and CD8 molecules, improving the jejunum structure. Meanwhile, B.subtilis increased the concentration of plasma AST and CD8 molecules of broilers. Additionally, B.subtilis also increases the plasma GSH-Px content, but not affecting the plasma biochemical indexes. And increases the plasma IgG and GSH-Px contents and the VCR ratio also improved in M-group. Additionally, the methionine content in thoracic muscle was increased in all groups. But only adding probiotics groups increase the relative abundance of broiler intestinal phylum Synechococcus and Phylum Firmicutes while decreasing the relative abundance of Enterococcus spp. Our results showed that adding probiotics could improve the meat output and quality. It might also be necessary to optimize the feeding conditions by selecting specific strains according to different effects of those probiotics, and to evaluate the best viable number of probiotics which survives in broiler guts to balance the cost and the effort.